Immunology Applications Project
Immunology Applications Project Bio 152 For this project, you will be working on internet-based programs that will walk you through some of the practical applications of immunology. These immunological assays can be used to detect a wide variety of diseases and are regularly used in doctor’s offices and hospitals. You can type your answers directly into this Word document; when you are done, print it out and turn it in on the day of the final exam. If you cut and paste from the website rather than putting things into your own words, you will not receive credit for the exercise. Start out by going to the following YouTube video http://www.youtube.com/watch?v=oXL1rH11MTg&feature=relmfu What is an agglutination assay? What is the basis of the way in which this assay work What types of antigens can be detected by an agglutination assay? What types of tests are used clinically that are based on agglutination assay Now, go to the following YouTube video to learn the basics about the ELISA. This will give you some background information for the next section of the exercise. http://www.youtube.com/watch?v=RRbuz3VQ100&feature=relmfu To get started on the next exercise, go to the following website: https://media.hhmi.org/biointeractive/vlabs/immunology/index.html 1 Once the website loads, you will see a page that has a picture on the left side of a laboratory, and a series of tabs on the right side that will allow you to work through the lab exercise. First, 1. read the directions on the Diagnosis tab on the right side of the page 2. read the background information that is on the second tab 3. click on the Notebook tab. You will work through this tab in conjunction with the interactive pictures on the left side of the page in order to complete the project. What does the term ELISA mean? Describe the four steps in the generalized ELISA protocol for detecting a target antigen. Why must the patient’s whole blood samples be centrifuged at the start of the laboratory assay? Why must you prepare three dilutions of the serum samples? What does the acronym PBS mean? Why have the wells of the ELISA plate been pre-treated with SLE antigen? 2 Why are both a positive and negative control used when running the test? What is used as the positive control? What is used as the negative control? What is meant by the terms primary antibody and secondary antibody? How are they different from each other? What does HRP stand for? What is its purpose in the test system? What is the purpose of incubating the ELISA plate at 37 degrees C for 15 minutes? What will happen if the plate is not incubated properly? What is the purpose of washing the wells in the plate with PBS after the incubation period? What is the secondary antibody that is used in this experiment? How is it different from the primary antibody? What is the enzyme that is attached to this secondary antibody? 3 What is the purpose of the second incubation in the process? Why is it incubated at 37 degrees C? What is the substrate for HRP in the test? How does the HRP work with the substrate to result in a yellow color if the test result is positive? What are the results of the tests on the three patients? Who has the disease? Are the results of the test unequivocal in each test? Print out the summary page of this exercise and attach it to this exercise when you turn it in. 4 Now, go to the following animation/ video to learn something about monoclonal antibody production. http://www.sumanasinc.com/webcontent/animations/content/monoclonalantibodies.h tml What is a monoclonal antibody? What is a hybridoma cell? Where does it come from? What is the purpose of this cell? 5
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